Bicistronic expression of nuclear transgenes in Chlamydomonas reinhardtii.
Chlamydomonas reinhardtii
bicistronic gene expression
microalga
nuclear transgene engineering
polycistron
transgene design
Journal
The Plant journal : for cell and molecular biology
ISSN: 1365-313X
Titre abrégé: Plant J
Pays: England
ID NLM: 9207397
Informations de publication
Date de publication:
28 Feb 2024
28 Feb 2024
Historique:
revised:
19
01
2024
received:
28
07
2023
accepted:
29
01
2024
medline:
28
2
2024
pubmed:
28
2
2024
entrez:
28
2
2024
Statut:
aheadofprint
Résumé
In eukaryotic organisms, proteins are typically translated from monocistronic messenger RNAs containing a single coding sequence (CDS). However, recent long transcript sequencing identified 87 nuclear polycistronic mRNAs in Chlamydomonas reinhardtii natively carrying multiple co-expressed CDSs. In this study, we investigated the dynamics of 22 short intergenic sequences derived from these native polycistronic loci by their application in genetic constructs for synthetic transgene expression. A promising candidate sequence was identified based on the quantification of transformation efficiency and expression strength of a fluorescence reporter protein. Subsequently, the expression of independent proteins from one mRNA was verified by cDNA amplification and protein molecular mass characterization. We demonstrated engineered bicistronic expression in vivo to drive successful co-expression of several terpene synthases with the selection marker aphVIII. Bicistronic transgene design resulted in significantly increased (E)-α-bisabolene production of 7.95 mg L
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : King Abdullah University of Science and Technology
Informations de copyright
© 2024 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.
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